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
methods used to classify them are not com-
pletely satifactory. Generally, the scheme de-
vised by Baird-Parker (2, 3) is employed to
divide the family Micrococcaceae. However,
with this scheme it is impossible to separate
weakly anaerobic staphylococci from micro-
cocci, and such strains are often misidentified
as micrococci. In applying Baird-Parker’s sys-
tem, Noble (12,13) has found that the predomi-
nant groups of skin cocci are Staphylococcus
type 2 (probably S. epidermidis sensu stricto)
and Micrococcus type 2. The taxonomic status
of the latter type, which may include some
weakly anaerobic staphylococci, is not clear.
Deoxyribonucleic acid (DNA) base composition
and cell wall composition are the characters
most; useful in separating Micrococcus from
Stuphylococcus (16, 18, 19). The cell wall pepti-
doglycan of the staphylococci contains large
amounts of glycine and is thus clearly different
from that of the micrococci. Moreover, all
staphylococci contain teichoic acids or similar
50
cell wall composition and the type of lactic acid
produced from glucose under anaerobic condi-
tions are very valuable in differentiating staphy-
lococci (20). This paper reports on the results
of the application of these criteria, as well as of
morphological and physiological characters, to
the classification of staphylococci isolated from
human skin.
MATERIALS AND METHODS
Bacterial strains. Staphylococci were isolated
from the healthy skins of two groups of people: 20
people living in Raleigh, N.C., who were sampled once
each month for 6 to 13 months, and 20 people living in
Somerville or New Brunswick, N.J., who were sam-
pled once during the winter. Samples were taken from
two separate sites on the forehead and one site from
one cheek, the chin, one external nare, anterior nare,
each axilla, each upper and lower arm, and each
upper and lower leg. Representative strains are listed
in Tables 2 through 5.
Isolation and culture techniques. The procedures
and media used for isolating staphylococci were
identical to those used in previous studies on micro-
cocci from human skin (10).
VOL.25, 1975
Morphological and colonial characters. Colony
characteristics and cell morphology were determined
as described previously for micrococci (10).
Physiological characters. Oxygen and tempera-
ture relationships and tolerance to NaCl were deter-
mined as described previously (10).
Biochemical characters. Catalase and benzidine
tests, DNA base composition, acetylmethylcarbinol
production, nitrate reduction, and carbohydrate reac-
tions were determined as described previously for
micrococci (10). Bound and free coagulase activities
were determined on human, rabbit, and bovine plasma
(1, 24). Hemolysis was detected on bovine, human,
sheep, and rabbit blood agar plates after 24, 48, and
72 h by using streak cultures. Deoxyribonuclease pro-
duction was determined on deoxyribonuclease test
agar (Difco) plates after 24 and 48 h by using streak
streak cultures. Deoxyribonuclease (DNase) produc-
tion was determined on deoxyribonuclease test agar
(Difco) plates after 24 and 48 h by using streak
cultures. Phosphatase activity was determined by a
modification of the technique of Pennock and Huddy
(14), using a 0.005 M phenolphthalein monophos-
phate sodium salt (Sigma) instead of a 0.01 M
solution of disodium phenylphosphate. Production of
bacteriolytic activity was detected by using freeze-
dried M . luteus cells (lysozyme substrate, Difco) (26)
or living M. luteus (ATCC 27141) cells in an agar
overlay.
Lysostaphin susceptibility. An agar overlay was
prepared by adding 0.1 ml of a saline cell suspension
(approximately 10' colony-forming units per ml) to a
tube containing 3 ml of fluid, soft peptone-yeast
extract-glucose agar (10). The contents were mixed
thoroughly and then poured on the surface of a dry
soft peptone-yeast extract-glucose agar plate. A drop
of sterile lysostaphin solution (200 pg/ml) was placed
on the inoculated agar and incubated for 24 to 48 h.
Lysostaphin susceptibility was interpreted according
to the following spot-inhibition scheme: + sensitive
(complete growth inhibition); *, slightly resistant
(partial growth inhibition); and -, resistant (no visi-
ble growth inhibition). The minimal inhibitory con-
centration of lysostaphin was determined on agar
plates containing different concentrations of lyso-
staphin. A loopful of a saline cell suspension was
placed on each plate. Plates were incubated for 48 h
and then examined for growth.
Lysozyme susceptibility. Lysozyme susceptibil-
ity was determined on the same agar plate as used for
testing lysostaphin. A drop of a sterile lysozyme
solution (400 pglml) was placed on the agar plate.
Lysozyme susceptibility was interpreted by using the
same scheme described for the lysostaphin suscepti-
bility test.
Antibiotic susceptibilities. Procedures for testing
susceptibility to penicillin G, streptomycin, erythro-
mycin, tetracycline, and- novobiocin were described
previously ( 10).
Cell wall preparation and analyses. Cell walls
were prepared and analyzed as described previously
(17, 20). The different peptidoglycan types are abbre-
viated by the system of Schleifer and Kandler (19).The
procedure of Wolin et al. (27) was used for qualitative
STAPHYLOCOCCI FROM HUMAN SKIN. I. 51
examination of the cell wall teichoic acids. In cases
where both glycerol and ribitol were found in the acid
hydrolysates, the teichoic acids were checked more
carefully. The cell walls were extracted with phenol
(80% [wt/vol] in water) or hot water to remove traces
of membrane teichoic acid. For some representative
strains of each group, the teichoic acid composition
was also studied more accurately by gas chromatogra-
phy or by enzymatic methods (20).
Determination of the type of lactic acid pro-
duced. The procedure for determining the type of
lactic acid produced was described previously (20).
Organisms were grown in yeast extract-peptone-
glucose broth under microaerophilic or anaerobic
conditions. They were incubated at 34 C for 5 to 6
days. The type of lactic acid produced was deter-
mined enzymatically with L-lactate dehydrogenase
from rabbit muscle (Boehringer, Mannheim) and
D-lactate dehydrogenase obtained from Leuconostoc
mesenteroides ATCC 12291 (6).
Chemicals. Egg white lysozyme (muramidase) was
purchased from Sigma Chemical Co., St. Louis. Mo.;
lysostaphin was from Schwarz/Mann, Orangeburg,
N.Y. T h e following carbohydrates were tested:
D( + )-glucose, D-D( - )-fructose, D( +)-galactose, D( + ) -
mannose, L(+)-rhamnose, D( +)-xylose, L ( + ) -
arabinose, D( - )-ribose, maltose, a-lactose, sucrose,
D( +)-trehalose, D( +)-turanose, P-gentiobiose, D( + )-
cellobiose, D( + )-melezitose, raffinose, D-mannitol,
glycerol, xylitol. D-sorbitol, rneso-inositol, salicin,
adonitol (ribitol), dulcitol, earabitol, rneso-erythritol,
D-erythrose. P-melibiose, a-L-fucose, tagatose, D-lyx-
ose, and L-sorbose.
RESULTS AND DISCUSSION
Identification of the genus Staphylococcus.
The main characteristics for the differentiation
of the genera Micrococcus and Staphylococcus
are listed in Table 1. Members of the genus
Staphylococcus possess a low guanine plus cyto-
sine ( G + C ) content (30 to 38 mol%) in their
DNA, whereas members of the genus
Micrococcus have a rather high G + C content
(66 to 73 mol%). These differences in the DNA
base composition are accompanied by differ-
ences in the chemical composition of the cell
walls, as discussed above. The difference in the
cell wall peptidoglycan composition is reflected
by the susceptibility lysostaphin. The main bac-
teriolytic activity of lysostaphin is due to an
endopeptidase with a specificity of action
against the pentaglycine interpeptide bridge of
the peptidoglycan. Thus, all staphylococci
tested by the spot test described above showed
some susceptibility to lysostaphin, whereas
most micrococci were resistant.
Only a few micrococci that were susceptible
to lysozyme showed a weak susceptibility to ly-
sostaphin. These strains, however, cannot be
confused with staphylococci, since all of the
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
about 30% showed weak hemolysis activity. All
strains were catalase and benzidine test posi-
tive. All produced acetylmethylcarbinol. More
than 80% of the strains reduced nitrate or dem-
onstrated phosphatase activity. About half of
the strains showed a weak extracellular deoxy-
ribonuclease activity. Two-thirds of the strains
demonstrated b act eriolyt ic activity .
All strains produced acid aerobically from
glucose, fructose, maltose, sucrose, and glyc-
erol. Seventy to 90% of the strains produced
acid from galactose, mannose, lactose, or turan-
ose. Less than 20% produced acid for ribose or
melezitose. No acid was produced from rham-
nose, xylose, arabinose, trehalose, gentiobiose,
cellobiose, mannitol, xylitol, sorbitol, inositol,
salicin, adonitol, dulcitol, arabitol, erythritol,
erythrose, raffinose, melibiose, fucose, tagatose,
lyxose, or sorbose.
All strains were either susceptible or slightly
resistant to lysostaphin and were resistant to
lysozyme. They were usually susceptible to
novobiocin. Most strains were susceptible to
penicillin, erythromycin, and tetracycline.
Most strains were resistant to streptomycin (5
pg/ml).
Twenty-one strains studied previously (group
IIAI, reference 20) and 20 strains isolated from
skin contained peptidoglycan of type L-LYS-
Glyr-a,L-Sero.7-l.5, The cell wall teichoic acid of
these strains was composed of glycerol and
glucose.
All strains lowered the pH of yeast extract-
glucose broth under anaerobic conditions from
6.8 to 4.3-4.6. They produced predominantly
L-lactic acid and only small amounts (usually
less than 10%)of D-lactic acid. The type strain,
ATCC 14990 (8), and 20 strains studied previ-
ously (group IIA1, reference 20) contained an
L-lactate dehydrogenase, which is activated by
fructose-176-diphosphate.
The G+C content of the DNA, as determined
in six strains, is 33.5 * 0.2 mol%.
Recent studies by Schleifer and Steber (21)
showed t h a t phages isolated from S .
epidermidis strains by Verhoef et al. (25) ad-
sorbed only to cell walls showing the character-
istic cell wall composition of s. epidermidis. In
particular, the presence of glucosylated glycerol
teichoic acid was necessary for specific adsorp-
tion.
S. epidermidis can be distinguished from
other staphylococci primarily by its cell wall
composition, colony morphology, growth on
NaCl agar, carbohydrate reaction pattern, and,
to a lesser extent, on the type of lactic acid
produced, growth in thioglycolate medium,
ph 0sp h a t as e activity, ac et y lm et h y lcar b inol
STAPHYLOCOCCI FROM HUMAN SKIN. I. 53
production, nitrate reduction, growth a t ex-
treme temperature, and susceptibility to novo-
biocin.
(ii) S. saprophyticus Fairbrother emend.
Shaw et al. 1951. The following description of S .
saprophyticus is based on a total of 83 strains,
unless noted otherwise.
Cells were gram-positive cocci with a mean
diameter of 0.8 to 1.2 pm; they were nonmotile,
nonsporeforming, and occurred predominantly
in pairs or as single cells.
Colonies were raised to slightly convex, circu-
lar, usually entire, 4.0 to 9.0 mm in diameter,
smooth, glistening, and usually opaque. Colony
pigment was variable; however, most strains
were unpigmented or had a slight yellow tint
which increased in intensity with age.
Growth occurred in both the aerobic and
anaerobic portions of thioglycolate medium;
growth in the anaerobic portion was usually
dense or showed a gradient from dense to light
growth in the deeper, more anaerobic, portion of
this medium. Seventeen of the strains isolated
from skin and strains CCM 883 ( = ATCC 15305
= NCTC 7292; type strain, reference 22)72204,
and P.O. 3519 (obtained from P. Oeding, Ber-
gen, Norway) lowered the pH in yeast extract-
glucose broth under anaerobic conditions from
6.8 to 5.0-5.4. Low amounts of lactic acid,
primarily L-lactic acid, were produced by most
strains (Table 2). All strains grew well or at
least poorly at concentrations of NaCl up to
15%. All strains grew well at 15 to 40 C; 36% of
the strains grew poorly and 26% failed to grow a t
45 c.
None of the strains produced coagulase or
extracellular deoxyribonuclease or showed bac-
teriolytic or hemolysin activity. All strains were
positive for catalase and the benzidine test. All
strains produced acetylmethylcarbinol and,
with the exception of strain SE 11, did not
reduce nitrate and did not demonstrate phos-
phatase activity.
All strains produced acid aerobically from
glucose, maltose, sucrose, turanose, mannitol,
and glycerol. At least 80% of the strains pro-
duced acid from fructose, lactose, trehalose, and
xylitol. Strain SE 11 produced acid from galac-
tose and strain KL 150 produced acid from
gentiobiose. No acid was produced from man-
nose, rhamnose, xylose, arabinose, ribose, cel-
lobiose, melezitose, sorbitol, inositol, salicin,
adonitol, dulcitol, arabitol, erythritol, ery-
t h o s e , raffinose, melibiose, fucose, tagatose,
lyxose, or sorbose.
All strains were susceptible to lysostaphin
and resistant to lysozyme. All strains were
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
erythromycin. All strains were resistant to novo-
biocin ( 5 pg/ml).
Seventeen strains isolated from skin and
strains CCM 883,2204, and P.O. 3519 contained
.
peptidoglycan of type ~ - L p - G l y , -,, L-Sero-6-o.8
Their cell wall teichoic acids contained two
polyols: glycerol and ribitol. In a previous study
(20), only ribitol was determined as a polyol
constituent of the cell wall teichoic acid of
strains CCM 883 and 2204, and related strains
(group IIB), but gas chromatographic and en-
zymatic analyses showed that glycerol, al-
though usually in much smaller amounts than
ribitol, is also present.
The G + C content of the DNA, as determined
in five strains, is 33.5 A 1 mol%.
Some variable characters and the G + C con-
tent of the DNA of representative strains of s.
suprophyticus are shown in Table 2.
S . saprophyticus can be distinguished from
other staphylococci primarily on the basis of cell
wall composition, carbohydrate reaction pat-
tern, and, to lesser extent, on low acid produc-
tion under anaerobic conditions, production of
acetylmethylcarbinol, the failure to reduce ni-
trate, and resistance to novobiocin.
(iii) S. cohnii sp.n. (c0h'ni.i. M.L. gen. noun
of Cohn; named for Ferdinand Cohn, a German
botanist and bacteriologist). The following de-
scription of S . cohnii is based on a total of 42
strains, unless noted otherwise.
Cells were gram-positive cocci, 0.5 to 1.2 pm
in diameter, occurring predominantly singly or
in pairs (tetrads were found only in a few
strains). Nonmotile and nonsporeforming.
Colonies were convex to slightly peaked (a
few strains had colonies with depressed cen-
ters), flat edged, circular, 4.0 to 7.5 mm in
diameter, entire, smooth, usually glistening or
glossy, opaque, and usually unpigmented.
Growth occurred in both the aerobic and
VOL.25, 1975
anaerobic portions of thioglycolate medium;
growth in the anaerobic portion was either
uniformly dense or showed a gradient from
dense to light growth in the more anaerobic
portion of the medium. Twenty-four percent of
the strains grew only as single colonies in the
anaerobic portion.
Twenty-two strains isolated from skin did not
produce a pH lower than 5.3 in yeast extract-
glucose broth under anaerobic conditions. A
small amount of lactic acid was produced,
predominantly L-lactic acid, but some strains
produced DL-lactic acid.
All strains grew well in media containing up
to 10% NaC1, but 64% of the strains grew poorly
and 15%failed to grow in 15% NaC1. All strains
grew well a t 15 C, and most grew a t 45 C; only
7% failed to grow a t 45 C.
All strains failed to produce coagulase. All
strains were catalase and benzidine test posi-
tive. Thirty-one percent of the strains demon-
strated weak hemolysis activity and 55%
showed no hemolysis activity. More than 80% of
the strains produced weak to moderate acetyl-
methylcarbinol reactions and failed to reduce
nitrate. About 20% of the strains demonstrated
weak phosphatase and extracellular deoxyribo-
nuclease activities, and 38% exhibited bacteri-
olytic activity.
All strains produced acid aerobically from
glucose, fructose, trehalose, and glycerol. About
80% of the strains produced acid from mannose,
maltose, and mannitol. Some strains produced
acid from lactose (31%) or xylitol (38%).Very
few strains produced acid from sucrose (12%) or
galactose (10%). No acid was produced from
rhamnose, xylose, arabinose, ribose, turanose,
gentiobiose, cellobiose, melezitose, sorbitol,
inositol, salicin, adonitol, dulcitol, arabitol,
erythritol, erythrose, raffinose, melibiose, fu-
cose, tagatose, lyxose, or sorbose.
All strains were susceptible to lysostaphin
and resistant to lysozyme. About one-half of the
strains were resistant to streptomycin or eryth-
romycin. More than 90% of the strains were
susceptible to penicillin or tetracycline. About
90% of the strains were resistant to novobiocin
(5 pg/ml).
Twenty strains isolated from skin contained
peptidoglycan of type ~-Lys-Gly,.,. Their cell
wall teichoic acid contained glycerol and usu-
ally glucose; glucosamine was also present in
extracted teichoic acids, but generally in low
amounts. Some strains (KH 201, RM 10, JRM
24, JL 143, and BB 3) also contain galactosa-
mine in their cell walls.
The G+C content of the DNA, as determined
STAPHYLOCOCCI FROM HUMAN SKIN. I. 55
in three strains, is 37.2 A 0.5 mol%.
Variable characters of 20 selected strains and
the G + C contents of some representative
strains are listed in Table 3. Previously studied
strains (group IC 3, reference 19) containing a
similar cell wall composition may also belong to
this species.
Strain DSM 20260 (originally designated GH
137) is the type strain of S . cohnii. A description
of this strain follows.
Cells are gram-positive spheres, about 1.2 pm
in diameter, occurring predominantly in pairs
and singly, occasionally in tetrads. Nonmotile
and nonsporeforming.
Agar colonies are circular, entire, about 6.0
mm in diameter, convex to slightly peaked with
a flat edge, smooth with glossy surface, white (a
color photograph of a colony is shown in Fig. lD,
reference 9), and opaque.
Chemoorganotrophic: Metabolism is respira-
tory and fermentative.
Catalase and benzidine test positive.
Weakly facultatively anaerobic.
The anaerobic portion of thioglycolate me-
dium showed a gradient from dense to light
growth in the deeper, more anaerobic, portion of
the medium. The pH of yeast extract-glucose
broth was lowered under anaerobic conditions
from 6.8 to 5.3. Small amounts of L-lactic acid
were produced.
Growth on NaCl agar: Good growth with 10%
NaC1, weak growth a t 15%.
Temperature relationships: Good growth a t
15 and 45 C.
Coagulase, phosphatase, extracellular deoxy-
ribonuclease, and bacteriolytic activities not
demonstrated.
Produced partial hemolysis of bovine blood
but not sheep blood.
Acetylmethylcarbinol produced.
Nitrates not reduced.
Acid produced aerobically from glucose, fruc-
tose, maltose, trehalose, mannitol, and glycerol.
No acid from galactose, mannose, rhamnose,
xylose, arabinose, ribose, lactose, sucrose, tu-
ranose, gentiobiose, cellobiose, melezitose,
xylitol, sorbitol, inositol, salicin, adonitol, dul-
citol, arabitol, erythritol, erythrose, raffinose,
melibiose, fucose, t agatose, lyxose, or sorbose.
Susceptible to lysostaphin (minimal inhibi-
tory concentration, 50 pg/ml); resistant to lyso-
zyme.
Antibiotic susceptibilities: Susceptible to
penicillin and tetracycline.
Resistant to streptomycin (5 pg/ml), erythro-
mycin (150 pg/ml), and novobiocin (20 pg/ml).
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
amounts.
G+C content of the DNA: 37.1 mol%.
S . cohrzii can be distinguished from all other
staphylococci primarily on the basis of carbohy-
drate reaction pattern, cell wall composition,
and, to a lesser extent, low acid production
under anaerobic conditions, production of ace-
tylmethylcarbinol, lack of nitrate reduction,
and resistance to novobiocin. There are also
some differences between S. cohrzii and other
staphylococcal species in colony pigment, cell
arrangement, growth a t extreme temperatures,
and coagulase, hemolysin, phosphatase, extra-
cellular deoxyribonuclease, and bacteriolytic
activities.
(iv) S. huemolyticus sp.n. (hae.mo.ly’ti.cus.
M.L.adj. huemolyticus blood-dissolving) . Sev-
eral strains of this species were isolated previ-
ously but were identified as belonging to Micro-
coccus halodurans, Micrococcus s ~ . ,or
epidermidis (5, 15; P . Oeding, personal commu-
nication). These strains include CCM 1798,
2444, and 2574; P.O. 375 (P. Oeding, Bergen,
Norway); and S 1and S 226 (S. Schaefler, New
York, N.Y.). Based on their cell wall composi-
tions and lactic acid configurations, these and
other strains were placed in staphylococci group
IIA4 (20). The following description of S.
huemolyticus is based on a total of 117 strains,
Cells were gram-positive cocci, 0.8 to 1.3 pm
in diameter, occurring predominantly in pairs
and tetrads. Nonmotile and nonsporeforming.
Colonies were raised to slightly convex, circu-
lar, 4.0 to 9.0 mm in diameter, usually entire,
smooth, glistening, and usually opaque. Colony
pigment was variable; however, most strains
were unpigmented or had a slight yellow tint
which increased in intensity with age.
Growth occurred in both the aerobic and
anaerobic portions of thioglycolate medium. In
the anaerobic portion, there was a gradient from
dense to light growth or individual colonies in
the deeper, more anaerobic, portion of the
medium.
Twenty-three strains isolated from skin and
strains CCM 1798, 2444, and 2574, P.O. 375, S
1, and S 226 fermented glucose under anaerobic
conditions and lowered the pH of yeast extract-
glucose broth from 6.8 to 4.9-5.5. These strains
s. usually produced D-lactic acid from glucose; two
strains also produced 15 to 30% L-lactic acid.
All the strains grew well at the NaCl concen-
trations tested up to 10%; 40% of the strains did
not grow with 15%NaCl. All strains grew well at
45 C, but 57% of the strains failed to grow a t
15 C.
All strains were catalase and benzidine test
positive. All failed to produce coagulase. All
strains showed moderate or weak hemolysis.
VOL. 25, 1975
The hemolysis was best demonstrated with
bovine blood and is slightly less intense with
sheep and rabbit blood. More than 80% of the
strains reduced nitrate and produced acetyl-
methylcarbinol. Most strains failed to demon-
strate extracellular deoxyribonuclease and
phosphatase activity; only 20 to 30% of the
strains showed weak activities. Bacteriolytic
activity was variable.
All strains produced acid aerobically from
glucose, maltose, sucrose, trehalose, and glyc-
erol. About one-half of the strains produced acid
from fructose, galactose, lactose, turanose, and
mannitol. Only a few strains produced acid
from mannose (3%)and ribose (17%).All strains
failed to produce acid from rhamnose, xylose,
arabinose, gentiobiose, cellobiose, melezitose,
xylitol, sorbitol, inositol, salicin, adonitol, dul-
citol, arabitol, erythritol, erythrose, raffinose,
melibiose, fucose, tagatose, lyxose, or sorbose.
All strains were slightly resistant or suscepti-
ble to lysostaphin and resistant to lysozyme. All
strains, except S 1 and S 226, were susceptible
to novobiocin and 90% of the strains were
susceptible to erythromycin. Most were suscep-
tible to penicillin (79%)and tetracycline (91%).
Twenty-three strains isolated from skin and
strains CCM 1798, 2444, and 2574, P.O. 375, S
1, and S 226 contained the cell wall peptidogly-
can of type L - L ~ S - G ~ ~ ~ . ,L-Sero.,-l.,.
,.,, Their
cell wall teichoic acid was composed of glycerol
and glucosamine.
T h e G + C content of DNA, as determined for
three strains, was 35.3 f 0.6 mol%.
Some variable characters of 23 selected
strains from skin and the G+C contents of the
DNA of representative strains are given in
Table 4.
Strain DSM 20263 (originally designated SM
131) is the type strain of S . huemolyticus. A
description of this strain follows.
Cells were gram-positive spheres, about 1.3
pm in diameter, occurring predominantly in
tetrads and less frequently in pairs. Nonmotile
and nonsporeforming.
Agar colonies were circular, entire, 5 to 6 mm
in diameter, slightly convex. White with a
yellowish tint (see color photograph of colony:
Fig. lF, bottom row, reference 9), and smooth
with glistening surface.
Chemoorganotrophic: Metabolism is respira-
tory and fermentative.
Catalase and benzidine tests were positive.
Facultatively anaerobic.
Growth in the anaerobic portion of thioglyco-
late medium occurred only in individual colo-
nies.
The pH of yeast extract-glucose broth was
lowered under anaerobic conditions from 6.8 to
STAPHYLOCOCCI FROM HUMAN SKIN.I. 57
5.0. Lactic acid was produced solely as the D
isomer.
Growth on NaCl agar: Good growth a t lo%,
no growth a t 15% NaCl.
Temperature relationships: No growth a t
15 C, good growth at 45 C.
Bovine and sheep erythrocytes were hemo-
lyzed.
Coagulase, phosphatase, extracellular deoxy-
ribonuclease, and bacteriolytic activities were
not demonstrated.
Acetylmethylcarbinol not produced.
Nitrates were reduced.
Acid was produced from glucose, maltose,
sucrose, trehalose, and glycerol. No acid from
fructose, galactose, mannose, rhamnose, xylose,
arabinose, ribose, lactose, turanose, gentiobi-
we, cellobiose, melezitose, mannitol, xylitol,
sorbitol, inositol, salicin, adonitol, dulcitol, ara-
bitol, erythritol, erythrose, raffinose, melibiose,
fucose, tagatose, lyxose, or sorbose.
Susceptible to lysostaphin (minimal inhibi-
tory concentration, 100 pg/ml); resistant to
lysozyme.
Antibiotic susceptibilities: Susceptible to
penicillin, erythromycin, tetracycline, and no-
vobiocin. Resistant to streptomycin (5 pg/ml).
Cell wall peptidoglycan type: L-LYS-G~Y,.,,
L-Ser 2 .
Cell wall teichoic acid composition: Glycerol
and glucosamine.
G + C content of the DNA: 36.4 mol%.
S. huemolyticus can be distinguished from all
other staphylococci primarily by its hemolytic
activity, anaerobic growth pattern in thioglyco-
late medium, formation of D-lactic acid, and
cell wall composition. There are also some
significant differences between S . huemolyticus
and certain other species in colony pigment,
growth a t extreme temperatures, carbohydrate
reaction pattern, nitrate reduction, and the lack
of coagulase, phosphatase, extracellular deoxy-
ri bonuclease, and bacteriolyt ic activities.
(v) S. xylosus sp.n. (xy.lo’sus. M.L. adj.
xylosus xylose.) The following description of S.
xylosus is based on a total of 102 strains, unless
noted otherwise.
Cells were gram-positive cocci, 0.8 to 1.2 pm
in diameter. They were nonmotile and non-
sporeforming and occurred predominantly in
pairs or as single cells, occasionally as tetrads.
The colony morphology was highly variable.
Colonies were circular, 5.0 to 10.0 mm in
diameter, raised, slightly convex with a flat
edge or umbonate; the edges were entire, undu-
late, or crenate; the texture was smooth to
rough; and they were glistening to dull; they
were usually opaque. Colony pigment was also
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
yellow-gray. Some strains were highly sectored
with clones of different pigment intensities or
with pigmented and unpigmented clones. Some
strains were unpigmented a t 35 to 37 C but
developed intense pigment a t temperatures
below 20 C. Approximately 20% of the strains
were unpigmented a t all temperatures tested
from 15 to 45 C.
Growth occurred in both the aerobic and
anaerobic port ions of thioglycolate medium.
Growth in the anaerobic portion was often
dense; however, 25% of the strains demon-
strated less growth or produced only a few
colonies in the deeper, more anaerobic, portion
of this medium.
Twenty of the strains isolated from skin in
this study and strains CCM 1400 and 2210
lowered the pH of yeast extract-glucose broth
under anaerobic conditions from 6.8 to 4.9-5.6.
These strains produced predominantly L-lactic
acid.
All strains grew well a t NaCl concentrations
up to 10%; 48% of the strains grew poorly and
14% failed to grow a t a concentration of 15%.All
strains grew well at 15 C, but 62% failed to grow
a t 45 C.
None of the strains produced coagulase. All
strains were positive for catalase and the benzi-
dine test.
About 80% of the strains reduced nitrate and
failed to produce or produced only very low
levels of acetylmet hylcarbinol. All strains failed
to demonstrate hemolysis activity, and 80% of
the strains did not show an extracellular deoxy-
ribonuclease activity. About 80% of the strains
showed phosphatase activity. Bacteriolytic ac-
tivity was usually weak or absent.
All strains produced acid aerobically from
glucose, fructose, mannose, maltose, xylose,
mannitol, and glycerol. About 80% of the strains
produced acid from galactose, arabinose, lac-
tose, sucrose, trehalose, or turanose. Less than
30% of the strains produced acid from rham-
nose, ribose, gentiobiose, xylitol, sorbitol, or
inositol. None was produced from cellobiose,
melezitose, salicin, adonitol, dulcitol, arabitol,
erythritol, erythrose, raffinose, melibiose, fu-
cose, tagatose, lyxose, or sorbose.
All strains were susceptible to lysostaphin
and resistant to lysozyme.
All strains were susceptible to penicillin and
streptomycin. At least 80% of the strains were
susceptible to erythromycin and tetracycline.
About 70% were resistant to novobiocin (5
pdml).
Twenty of the strains isolated from skin in
this study and strains CCM 1400 and 2210 and
STAPHYLOCOCCI FROM HUMAN SKIN. I. 59
other related strains studied previously (group
IB, reference 20) contained peptidoglycan of
type ~-Lys-Gly,-,.The cell wall teichoic acid of
these strains contained glucosamine and, like
that of S. suprophyticus, two polyols: glycerol
and ribitol. Studies on strains CCM 1400 and
2606 proved that two distinct cell wall teichoic
acids, a ribitol and a glycerol teichoic acid,
occurred in these strains (Schleifer and van
Schaik, unpublished results).
The G + C content of the DNA, as determined
in six strains, is 34.2 f 1.1 mol%.
Some variable characters and the G+C con-
tent of DNA of some representative strains are
given in Table 5 .
Strains with similar properties were described
by Shaw et al. (22) as S. lactis. However, NCTC
7564 ( = ATCC 15306 = CCM 884) was desig-
nated by Shaw et al. as the type strain of S.
lactis, and this strain, a typical micrococcus
(18), was subsequently designated (11) as the
neotype strain of Micrococcus uuriuns Migula
1900. Thus, S. lactk is a later objective syn-
onym of M . uarians. Some of the strains previ-
ously identified as belonging to S. lactis are now
placed in S. xylosus.
Strain DSM 20266 (originally designated KL
162) is the type strain of S. xylosus. A descrip-
tion of this strain is presented below.
Cells were gram-positive cocci, about 1.2 pm
in diameter, occurring singly and in pairs. Non-
motile and nonsporeforming.
Agar colonies were circular, entire, about 4
mm in diameter, raised to slightly convex,
yellow-orange (see color photograph of a colony:
Fig. lC, middle row, reference 9), and opaque.
Chemoorganotrophic: Metabolism is respi-
ratory and fermentative.
Catalase and benzidine test positive.
Faculatively anaerobic.
Growth on NaCl agar: Good growth a t 10%
NaC1, poor growth a t 15% NaC1.
Temperature relationships: Good growth a t
15 C, but no growth a t 45 C.
Coagulase not produced.
Nitrates were reduced.
Acetylmethylcarbinol not produced.
Acid produced from glucose, fructose, galac-
tose, mannose, xylose, arabinose, maltose, lac-
tose, sucrose, trehalose, turanose, mannitol,
and glycerol.
Acid not produced from rhamnose, ribose,
cellobiose, melezitose, xylitol, sorbitol, inositol,
salicin, adonitol, dulcitol, arabitol, erythritol,
erythrose, raffinose, melibiose, fucose, tagatose,
lyxose, or sorbose. Yeast extract-glucose broth is
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.
Susceptible to lysostaphin (minimal inhibi-
tory concentration, 50 pg/ml) and resistant to
lysozyme.
Antibiotic susceptibilities: Susceptible to
penicillin, erythromycin, streptomycin, and tet-
racycline. Resistant to novobiocin ( 5 pg/ml).
Cell wall peptidoglycan type: L - L Y S - G ~Cell
Y~.
wall teichoic acid composition: glycerol, ribi-
tol, and glucosamine.
G+C content of the DNA: 36.2 mol%.
S . xylosus can be distinguished from all the
other staphylococci primarily on the basis of
its carbohydrate fermentation pattern, cell
wall composition, and growth a t extreme tem-
peratures and, to a lesser extent, on its reduc-
tion of nitrate, production of phosphatase, re-
sistance to novobiocin, and low acid production
under anaerobic conditions.
Several of the properties overlap with those of
S . cohnii and S. saprophyticus, suggesting a
relatively close relationship among these three
species. With respect to cell wall composition,
S. xylosus and S . cohnii contain identical pepti-
doglycan types, whereas with S. xylosus and S .
saprophyticus the chemical compositions of the
cell wall teichoic acids are identical. Low acid
production from glucose under anaerobic condi-
tions and resistance to novobiocin are properties
common to all three species. Moreover, the
three species have a lower optimal growth
temperature range than most other staphylo-
cocci. The common properties of the novobi-
ocin-resistant species may have been the reason
why the strains of these species were classified
in the new edition of Bergey's Manual (Baird-
Parker, personal communication) as belonging
to S . saprophyticus. Our studies, however, have
clearly demonstrated that a separation of these
novobiocin-resistant strains into three distinct
species is feasible. The cell wall peptidoglycan
type differentiates S. saprophyticus from S .
cohnii and S. xylosus, and the chemical compo-
sition of the cell wall teichoic acids separates S .
cohnii from S . xylosus and S . saprophyticus. In
addition, the failure to reduce nitrate and the
lack of phosphatase activity distinguish S.
saprophyticus and S . cohnii from most strains
of S. xylosus. The carbohydrate reaction pat-
tern is also a valuable character for the separa-
tion of these three species. Thus, S . xylosus
strains usually ferment xylose, arabinose, and
8-gentiobiose in contrast to most other staphy-
lococci. Most strains of S . saprophyticus are
distinguished by their ability to ferment xylitol,
whereas S. cohnii strains usually differ from
other staphylococci by their inability to ferment
sucrose or turanose.
ACKNOWLEDGMENTS
We are greatly indebted to E. Hagner, M. Musselwhite, I.
Pomper, and B. Popp for their excellent technical assistance
Va. 25, 1975
throughout this study.
This research was supported by a grant of the Deutsche
Forschungsgemeinschaft and by Public Health Service re-
search grant AI 08255 from the National Institute of Allergy
and Infectious Diseases.
REPRINT REQUESTS
Address requests for reprints to: Dr. K. H. Schleifer,
Lehrstuhl Mikrobiologie, Universitat Munchen, 8 Miinchen
19,Menzinger Str. 67,BRD, Germany.
LITERATURE CITED
1. Bailey, W. R., and E. G. Scott. 1966.Diagnostic microbi-
ology. The c. V. Mmby Co., St. Louis.
2. Baird-Parker, A. C. 1963.A classification of micrococci
and staphylococci based on physiological and biochem-
ical tests. J. Gen. Microbiol. 30:409-427.
3. Baird-Parker, A. C. 1965.The classification of staphylo-
cocci and micrococci from world-wide sources. J. Gen.
Microbiol. 38:363-387.
4. Bohacek, J., M. Kocur, and T. Martinec. 1965.Deoxyri-
bonucleic acid base composition and taxonomy of the
genus Micrococcus. Publ. Fac. Sci. Univ. J.E. PurkynB,
Bmo. K35:318-322.
5. Buck, J. D. 1965.Micrococcus halodumna nsp. Int. Bull.
Bacteriol. Nomencl. Taxon. 15:181-184.
6. Dennis, D. 1962. Lactic acid racemase from Clostridium
butylicum, p. 430-432. In S. P. Colowick and N. 0.
Kaplan (ed.),Methods in enzymology, vol. 5. Aca-
demic Press Inc., New York.
7. Evans, J. B., and W. E. Kloos. 1972.Use of shake cultures
in a semisolid thioglycolate medium for differentiating
staphylococci from micrococci. Appl. Microbiol.
23~326-331.
8. Hugh, R., and Me. A. Ellis. 1968.The neotype strain for
Staphylococcus epidermidis (Winslow and Winslow
1808) Evans 1916. Int. J. Syst. Bacteriol. 18:231-239.
9. Klooe. W. E.. and K. H. Schleifer. 1975. Isolation and
characterization of staphylococci from human skin. 11.
Deecription of four new species: Staphylococcus
warnerr, Staphylococcus capitia, Staphylococcus
hominis, and Staphylococcus simuhns. Int. J. Syst.
Bacteriol. 25:62-79.
10. K l m , W. E., T. G. Tornabene, andK. H. Schieifer. 1974.
Isolation and characterization of micrococci from
human skin, including two new species: Micrococcus
1yloe.and Micrococcus kristinae. Int. J. Syst. Bacteriol.
24:79-101.
11. Kocur, M., and T. Martinec. 1972.Taxonomic status of
Micrococcus varians Migula 1900 and designation of
the neotype strain. Int. J. Syst. Bacteriol. 22:228-232.
12. Noble, W. C. 1969.Skin carriage of the Micrococcaceae.
STAF'HnOCOCCI FROM HUMAN SKIN. I. 61
J . Clin. Pathol. 22:249-253.
13. Noble, W. C. 1969.Distribution of the Micrococcaceae.
Br. J. Dermatol. 81(Suppl. 1):27-32.
14. Pennock, C. A., and R. B. Huddy. 1967. Phosphatase
reaction of coagulase-negative staphylococci and mi-
crococci. J. Pathol. Bacteriol. 93:M-W.
15. Schaefler, S. 1971.Staphylococcus epidermidis BV: anti-
biotic resistance patterns, physiological characteris-
tics, and bacteriophage susceptibility. Appl. Microbiol,
22:693-699.
16. Schleifer, K. H.1973.Chemical composition of staphylo-
coccal cell walls, p. 13-32.In J. Jeljaszewicz and W.
Hryniewicz (ed.), Contributions to microbiology and
immunology, vol. 1. Staphylococci and staphylococcal
infections. S. Karger, Basel.
17. Schleifer, K. H., and 0. Kandler. 1967.Zur chemischen
Zusammensetzung der Zellwand der Streptokokken. I.
Die Aminostiuresequenz des Mureins von Streptococ-
cus thermophilus und Streptococcus faecalis. Arch.
Mikrobiol. 57:335-364.
18. Schleifer, K. H. and 0.Kandler. 1970. Amino acid
sequence of the murein of P~ROCOCCUS and other
Micrococcaceae. J. Bacteriol. 103:387-392.
19. Schleifer, K. H.,and 0. Kandler. 1972.The peptidogly-
can types of bacterial cell walls and their taxonomic
implications. Bacteriol. Rev. 36:407-477.
20. Schleifer, K. H.,and M. Kocur. 1973. Classification of
staphylococci based on the chemical and biochemical
properties. Arch. Mikrobiol. 93:65-85.
21. Schleifer, K. H.,and J. Steber. 1974.Chemische Untersu-
chungen am Phagenrezeptor von Staphylococcus
epidermidis. Arch. Mikrobiol. 98:251-270.
22. bhaw, C.,J. M. Stitt, and S. T. Cowan. 1951.Staphylo-
cocci and their classification. J. Gen. Microbiol.
5:1010-1023.
23. Silvestri, L. G., and L. R. Hill. 1965.Agreement between
deoxyribonucleic acid base composition and taxometric
classification of gram-positive cocci. J. Bacteriol,
90 ~136-140.
24. Subcommittee on Taxonomy of Staphylococci and Micro-
cocci. 1965. Recommendations. Int. Bull. Bacteriol.
Nomencl. Taxon. 15:109-110.
25. Verhoef, J., C. P. A. van Boven, and K. C. Winkler. 1971.
Lysogeny in coagulase-negative staphylococci. J. Med.
Microbiol. 4:405-412.
26. Wadstrom, T., and K. Hisatsune. 1970. Bacteriolytic
enzymes from Staphylococcus aureua. Purification of
an endo-B-N-acetylglucosaminidase. Biochem. J.
120:725-734.
27. Wolin, M. J., A. R. Archibald. and J. Baddiley. 1966.
Changes in wall teichoic acid resulting from mutations
of Staphylococcus aureus. Nature (London)
209:484-486.